Method of making blackleg vaccine



Patented Oct. 14, 1924.

UNITED STATES OLIVER M. FRANKLIN, OF AMABILLO, TEXAS, ASSIGNOR TO THE KANSAS BLAOKLEG SERUM COMPANY, LING, 01! DENVER, COLORADO, A CORPORATION OF COLORADO.

METHOD OF MAKING BLACKLEG VACCINE Io Drawing.

To all whom it may concern:

Be it known that I, OLIVER M. F RANKLIN, a citizen of the United States, and residing at Amarillo, Potter County, State of Texas, have invented certain new and useful Imrovements in Methods of Making Blackleg z1.ccine, of which the following is a specification.

My invention relates to a method paring a vaccine or bacterin for use in comhating the disesase among. cattle commonly known as blackle symtomatic anthrax. This disease is profuced by the blackleg organism which is now scientifically termed Ulostfidz'um cha'u/vei and which is well known to be a very resistant spore forming organism.

The object of the invention is to provide a method of preparing a vaccine for the purpose mentioned by growing in a suitable media. a culture of blackleg organisms, otherwise known as symtomatic anthrax, killing the organisms, and treating the product so as to obtain the organisms in a form suitable for use as a vaccine or bacterin to be injected into the cattle and produce immunity from the blackleg disease. More specifically the object of my invention is to prepare a vaccine or bacterin by inoculating into a suitable media a virulent or non-attenuated culture of the blackleg organisms and after suitable growth killing the organisms b the addition of a formalin solution (forma dehyde in water) or other suitable reagent and preferably at the same time heating the culture for a suitable length of time to a temperature not exceeding 50 0., and

en recovering the organisms fornuseas a vaccine. A further object of my invention is to provide a suitable culture media in which the blackleg organism may be successfully grown.

To attain the foregoing objects I proceed in the following manner. A suitable culture media is first prepared. This is done by grinding a quantity of suitable meat substance, preferably hog liver, and extracting the same with water, after which it is cooked and the broth pressed out of the ulp. The usual amount of peptone and sa t are. then added. It has been found that hog liver bouillon is particularl useful since it forms a, liquid media in w ich the blackleg organisms will grow satisfactorily without the addition of any other meat pulp or growth of pre- Application filed February 10, 1922. Serial No. 585,662.

accelerator except starch as hereinafter pointed out.

After the media is prepared as described, I place therein a cloth sack, preferably made of ordinary muslin, which contains calcium carbonate and starch in suitable quantities. The starch acts to accelerate growth of the organisms and the calcium carbonate tends to stabilize or keep the hydrogen ion concentration near neutral and also apparently fa.- cilitates growth of the organisms. This feature of placing the calcium carbonate and starch in a cloth sack has been found to be of considerable utility since it prevents the substances in the sack from mixing with the media and thus simplifies the subsequent separation of the materials in the sack from the organisms in the media. The media thus prepared is then sterilized in the usual manner.

While the blackleg bacillus is considered to be strictly anaerobic, I have found that the organism can be satisfactorily grown under aerobic conditions in a media prepared in the manner heretofore described. In other words, in a liquid media of this kind, the blackleg bacillus can be grown without subjecting it to strict anaerobic conditions or without special anaerobic apparatus. This media therefore possesses particular utility in my process.

After the media has been prepared and sterilized as described, I inoculate the same with a purified culture of blackleg organisms. For such inoculation I employ a freshly isolated culture of blackle preferably one obtained from a natural y occurring case of blackleg disease. In other words, Iinoculate the media with a non-at tenuated, pathogenic or virulent culture of the blackleg organisms. After suitable growth of the organisms has taken place the media is strained through any suitable substance, such as cotton or fine wire gauze, which serves to remove therefrom the coarser solid particles. A formalin solution 13 then added to the liquid containing the culture, and for this purpose a solution of ordinary commercial strength, approximately 37 to 40% formaldehyde, is preferably employed. The amount of formalin solution used may vary from about $4 to 1% of the volume of the media, but preferably about is emloyed. The resulting product is then allbwed to stand for several days, about 3 to 7, and during this period it is preferably vaccine which consists in growing blackleg kept at a temperature of approximately organisms in a suitable culture media, 0. as by means of a water bath or in any treating the same after suitable growth with other desired manner. The treatment with a formalin solution in concentration sufli- 5 formalin results in killing the organisms in cient to kill the organisms without destroytheir vegetative and spore forms and this ing their antigenic properties, and treatresult is accomplished 1].] less time and more ing the resulting roduct to obtain the satisfactorily if heat is employed as deorganisms in suitab e form for immunizing scribed. The killing of the organisms does use.

10 not destroy their antigenic property, that is 3. The method of preparing a blackleg they retain the property of causing reacvaccine which consists in rowing blackle tions in the treated animal as the result of organisms in a suitable cu ture media, adtfi which it becomes more resistant or immune ing a reagent to kill the organisms without to the blackleg disease. destroying their antigenic properties, heat- 16 After the foregoing treatment, the culture ing the solution, and treating the resulting is allowed to settle, most of the organisms product to obtain the organisms in suitable settling out in a short time. The product is form for immunizing use. then treated in any suitable way to obtain 4. The method of preparing a blackleg the organisms in a form for use as a vaccine vaccine which consists in growing blackleg 20 or bacterin to be in'ected into the cattle and organisms in a suitable culture media, treatproduce immunity rom the blackleg disease. ing the same after suitable growth with a One method of procedure is to decant the formalin solution in concentration suflicient su ernatant liquid to about of its original to kill the organisms without destroying vo ume, the remaining liquid containing the their antigenic properties, heating the solu- 26 organisms beingl used as a vaccine. An altion, and treating the resultin product to ternative proce ure is to centrifu e the orobtain the organisms in suita is form for ganisms out of the media and ry them. immunizing use.

The dry powder thus obtained is used for 5. The method of preparing a blackleg direct injection as a vaccine by suspending vaccine which consists in growing blackl 30 the same in a suitable liquid suspension or, organisms in a suitable culture media, ad if desired, the dry powder can be made in ing a reagent to kill the organisms without the form of a pel et and used as a vaccine. destroying their antigenic properties, heat- I am aware that vaccines or similar subing the solution to a temperature not exstances for treating blackleg disease have ceeding 50 (1., and treating the resulting 85 heretofore been proposed, but in general product to obtain the organisms in suitable 190 such prior processes are 0 en to numerous form for immunizin use. objections, involving comp icated procedure 6. The method 0% preparing a blackleg and in some cases requiring the slaughter of vaccine which consists in owing blackle cattle. Furthermore, such processes result organisms in a suitable cufi ure media, ad

40 in vaccines which are more or less lacking ing a reagent to kill the organisms without 1 in uniformity, the result being that the destroying their antigenic properties, heattreatment of cattle with such vaccines has ing the solution for a period from 3 to 7 been attended with uncertainty, the injecdays to a temperature not exceeding 50 C. tion in some cases causing fatal proliferaand treating the resulting product to obtain 45 tion of the blackleg spores, killing the anithe organisms in suita le form for im- 11 mal, and in others resulting in no immunity munizing use. from the disease. The improved process 7 The method of preparing blackleg vacherein described is simple and highly eflicine which consists in growing blackhzg orcient and by thorough and extended tests it ganisms in a suitable culture media, a ding 50 has been found that the vaccines produced thereto a formalin solution of sufficient by such process are uniform in quality and strength to kill the organisms without deact in a ve satisfactory manner to produce stroymg their antigenic pro rties, heating immunity mm the blackleg disease. the solution for a period of mm 3 to 7 days Having thus described my invention what at a temperature not exceeding 50 0., and 55 I claim as new and desire to secure by treating the resulting roduct to obtain the Letters Patent is: organisms in suitable cm for immunizing 1. The method of preparing a blackleg use. vaccine which consists in rowing blackleg 8. The method of preparing a blackleg organisms in a suitable cu ture media, addvaccine, which consists in growing in a 60 ing a reagent to kill the organisms without suitable liquid media under aerobic condidestroyin their antigenic properties, and tions a culture of blackleg organisms, addtreating tie resulting product to obtain the ing to the culture after suitable growth organisms in suitable form for immunizing a reagent to kill the organisms without destroying their antigenic properties, and l 2. The method of preparing a blackleg treating t e resultant product to obtain the organisms in suitable form for immunizing use.

9. The method of preparing a blackleg vaccine which consists in owing blackleg organisms in a suitable cu ture media, adding thereto a reagent to kill the organisms without destroying their antigenic properties, and withdrawing a portion of the liquid'from the media while retaining the organisms therein, whereby a concentrated solution adapted for use as a vaccine is obtained.

10. A method of making a media for use in growing a bacterial culture which consists in preparing a suitable bouillon or meat extract, placing in the media a suitable quantity of ca cium carbonate and starch enclosed in a cloth container and sterilizing the media.

11. A method of makin a media for use in growing a culture 0 blackleg which consists in preparing a bouillon or extract of hog liver, placing in the media a suitable quantlty of calcium carbonate and starch enclosed in a cloth container and sterilizing 25 the media.

In testimony whereof I hereunto afix my signature.

OLIVER M. FRANKLIN.

DISCLAIMER 1,511,557.--0Zivr M. Franklin, Amarillo, Tex. METHOD or mum swim V'accms. Patent dated October 14, 1924. Disclaimer filed October 2, 1933,

by the assignee, The 0. M. Franklin Blaclcleg Serum Company. Hereb enters its disclaimer to claims numbered 10 and 11 'in said specification, which are in the following ords, to-witz 10. A method of making a media for use in growing a bacterial culture which consists in preparing a suite. le bouillon or meat extract, placing in the media a suitable quantity of calcium carbonate and starch enclosed in a cloth container and sterilizing the media.

11. A method of making a media for use in growing a culture of blackleg which consists preparing a bouillon or extract of ho liver, pl m the media a suitable quantity of calcium carbonate and starch en osed in a do container and sterilizing the media.

[Oficial Gazette October 81, 1933.] 

